The objectives of the research proposal are four-fold. 1) We will examine the relationship between the interacton of pyridoxal phosphate (Vitamine B6) with glucocorticoid receptors and transformation of receptor into a nuclear binding protein. Monoclonal anti-vitamin B6 antibodies will be employed to detect pyridoxal phosphate steroid receptor interaction. 2) We will utilize pyridoxal phosphate as a molecular probe to gain new information about the physico-chemical properties of glucocorticoid receptors. 3) We will investigate the potential usage of the anti-vitamin B6 analog 5-deoxypyridoxal as a regulator and modifier of glucocorticoid receptor structure and function in whole cell. 4) Finally, we propose to utilize (32P)pyridoxal phosphate to dual label glucocorticoid receptors and purify these (32P) pyridoxal phosphte-(3H)dexamethasone receptor complexes to homogeneity. The purification of receptors will involve use of two techniques, pyridoxal phosphate affinity chromatography and anti-pyridoxal phosphate antibodies which should simplify the task of steroid receptor purification. We will then characterize the interaction of pyridoxal phosphate with purified glucocorticoid receptors from a variety of tissue.